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Simple Summary: The fruit fly (Drosophila melanogaster) has been used as a model organism to study the olfactory system of insects thanks to the wide range of genetic tools available in this species. Among these tools, optogenetics allows the immediate alteration of the functioning of certain cells with light by the targeted expression of light receptor proteins in these cells. Thus, by successively expressing these receptors in different elements of the behavioral circuit, it is possible to evaluate their effect on the final behavior of the organism. However, the use of optogenetics to dissect the receptor elements of adult olfactory behavior presents a challenge because most odorants elicit gradual attraction or avoidance depending on their concentration, complicating the representative substitution of odor by light. In this work, we explore a dual excitation model in which the subject responds to various odorant concentrations while the olfactory receptor neurons are activated by light. The dose–response curve in these flies remains odorant concentration dependent, but with reduced sensitivity compared to olfactory stimulation alone. The existence of an effect associated with each of the two stimuli, odor and light, allows us to explore the quantitative contribution of the receptor elements to olfactory behavior also by optogenetics. Optogenetics enables the alteration of neural activity using genetically targeted expression of light activated proteins for studying behavioral circuits in several species including Drosophila. The main idea behind this approach is to replace the native behavioral stimulus by the light-induced electrical activation of different points of the circuit. Therefore, its effects on subsequent steps of the circuit or on the final behavior can be analyzed. However, the use of optogenetics to dissect the receptor elements of the adult olfactory behavior presents a challenge due to one additional factor: Most odorants elicit attraction or avoidance depending on their concentration; this complicates the representative replacement of odor activation of olfactory sensory neurons (OSNs) by light. Here, we explore a dual excitation model where the subject is responding to odors while the OSNs are optogenetically activated. Thereby, we can assess if and how the olfactory behavior is modified. We measure the effects of light excitation on the response to several odorant concentrations. The dose-response curve of these flies still depends on odor concentration but with reduced sensitivity compared to olfactory stimulation alone. These results are consistent with behavioral tests performed with a background odor and suggest an additive effect of light and odor excitation on OSNs. [ABSTRACT FROM AUTHOR]

• Incomplete genetic databases impact metabarcoding-based river biomonitoring. • About 21% of macroinvertebrate species in the Iberian Peninsula are not barcoded. • Sequencing local macroinvertebrates improved assignments and biodiversity estimates. • A complete checklist of Iberian river macroinvertebrate species is required. Water resources play a vital role in sustaining ecosystems, human life, food security, and biodiversity. However, these resources currently face serious threats caused primarily by human activities. Because of that, European Union have mandated that all its member states monitor the ecological status of their water bodies. One of the most commonly used bioindicator groups for routine river biomonitoring is benthic macroinvertebrates, which rely mainly on morphological identification at the family level and the use of diverse indices to estimate the ecological status of the river ecosystem. The morphological approach has certain limitations, and metabarcoding is now considered an alternative technique for biodiversity studies and biomonitoring of surface waters. However, the efficacy of this approach depends on the comparison of sequencing data with genetic databases, which are often incomplete and geographically biased. In this study, we collected 92 macroinvertebrate samples from rivers in the northern Iberian Peninsula to identify all possible species and generate new DNA sequences for the COI-5P region. The objectives were to perform region-specific enrichment of reference databases for the geographical area of northern Spain and to assess the impact, before and after this strategy, on the taxonomic assignments used for ecological status inference in metabarcoding samples. The results showed that 21% of morphospecies lacked reference sequences in the BOLD or GenBank databases, indicating that a significant number of macroinvertebrate species in the northwestern Iberian Peninsula lack reference sequences in major genetic databases. Enriching the BOLD databases with new sequences led to the detection of an average of 17.25 more BINs (species-assimilable gene cluster) and 1.54 more IBMWP taxa (river ecological status indicator macroinvertebrate taxa) per sample, causing changes in the ecological status determination for 16% of the samples, which clearly shows that improving the databases with local species sequences could enhance the detection and identification of new IBMWP taxa, potentially impacting ecological status inference from molecular data. Additionally, a historical analysis of Iberian macroinvertebrate addition in the BOLD database using public BOLD records revealed an asymptotic trend that can be partially mitigated by region-specific enrichment. These results indicate that a local biota sequencing effort can considerably improve metabarcoding results rather than waiting for a gradual improvement of genetic databases. In conclusion, we advocate for a creation an Iberian freshwater macroinvertebrate checklist and then the initiation of a coordinated effort for sequencing any unsequenced species from the region. [ABSTRACT FROM AUTHOR]

Simple Summary: The artisanal octopus fishery is a great tradition and has a large economic impact in Northern Spain. Despite its importance, there is scarce genetic information for this exploited population. The uniqueness of an exploited population and the effects that exploitation may have on its long-term survival is essential information that can be obtained by studying the past and current spatial and temporal genetic patterns. The mitochondrial DNA is used here as a genetic tool to gain insight into Northeast Atlantic octopus populations and their evolutionary history. Although it is not sensitive enough to detect differences in nearby populations, it allows us to differentiate the two main lineages that originated during the Quaternary glaciations. One of the lineages is present exclusively in northern latitudes and the other mainly in the south, although, for the first time, its presence is demonstrated also in the north of the Iberian Peninsula. This implies a more continuous distribution than previously thought for this lineage. We also detect temporal changes in the distribution of the two lineages in contact zones. These changes seem to be associated with the effect of changing oceanographic conditions. Future studies on these associations could be of interest for fisheries in the scenario of global climate change. Octopus vulgaris is one of the most harvested octopus species in the world. In the Iberian Peninsula, there are several small-scale fisheries that have a long-term tradition of harvesting octopus. The Asturias fleet (in Northern Spain) has an internationally recognized MSC label for its exploitation. Of concern, genetic assessments of exploited stocks are currently scarce, which could prevent the implementation of adequate managing strategies. We use two mitochondrial regions (cytochrome oxidase subunit 1 and control region) to analyze the genetic status and evolutionary events that conditioned octopus populations' characteristics in the Northeastern Atlantic. A total of 90 individuals were sampled from three different localities in the Iberian Peninsula as well as a location in Macaronesia. Temporal genetic analyses on Asturias and Algarve populations were also performed. Results indicated the absence of fine spatial genetic structuring but showed the Canary Islands (in Macaronesia) as the most distinct population. Our analyses detected two distinct clades, already described in the literature, but, for the first time, we confirmed the presence of the α-southern haplogroup in the Northern Iberian Peninsula. This result indicates a more continuous cline for the distribution of these two haplogroups than previously reported. Temporal changes in the distribution of both haplogroups in contact zones were also detected. [ABSTRACT FROM AUTHOR]